Volume 6, Number 1—February 2000
Carbapenem-Hydrolyzing Metallo-ß-Lactamase from a Nosocomial Isolate of Pseudomonas aeruginosa in France
Highlight and copy the desired format.
|EID||Poirel L, Collet L, Nordmann P. Carbapenem-Hydrolyzing Metallo-ß-Lactamase from a Nosocomial Isolate of Pseudomonas aeruginosa in France. Emerg Infect Dis. 2000;6(1):84-85. https://dx.doi.org/10.3201/eid0601.000117|
|AMA||Poirel L, Collet L, Nordmann P. Carbapenem-Hydrolyzing Metallo-ß-Lactamase from a Nosocomial Isolate of Pseudomonas aeruginosa in France. Emerging Infectious Diseases. 2000;6(1):84-85. doi:10.3201/eid0601.000117.|
|APA||Poirel, L., Collet, L., & Nordmann, P. (2000). Carbapenem-Hydrolyzing Metallo-ß-Lactamase from a Nosocomial Isolate of Pseudomonas aeruginosa in France. Emerging Infectious Diseases, 6(1), 84-85. https://dx.doi.org/10.3201/eid0601.000117.|
To the Editor: The carbapenems (meropenem and imipenem), the ß-lactams with the broadest spectrum, are stable to most ß-lactamases (1). Therefore, they are often used as antibiotics of last resort for treating nosocomial infections due to gram-negative bacteria resistant to other ß-lactams. Resistance to carbapenems and susceptibility to other ß-lactams in Pseudomonas aeruginosa is common as a result of reduced drug accumulation or increased expression of pump efflux (1).
Several extended-spectrum ß-lactamases have been reported in P. aeruginosa, but only two, IMP-1 and VIM-1, possess an extended hydrolysis profile that includes carbapenems (2-5). The chromosome-borne and plasmid-mediated carbapenem-hydrolyzing ß-lactamase, IMP-1, has been described in several gram-negative rods, including P. aeruginosa, P. cepacia, Alcaligenes xylosoxydans, and Enterobacteriaceae isolates in Japan (4,6). Recently, a chromosome-borne carbapenem-hydrolyzing ß-lactamase, VIM-1, was reported from a clinical isolate of P. aeruginosa in Italy (5), and uncharacterized carbapenem-hydrolyzing ß-lactamases have been reported in the United Kingdom and Portugal (7,8). The weakly related IMP-1 and VIM-1 (31.4% amino acid identity) are both zinc-dependent (metallo-enzymes) and confer resistance to all ß-lactams except monobactams (3,5).
In 1996, a 39-year-old French woman was hospitalized in Marseille for chronic myelogenous leukemia, pancytopenia, and allogeneic bone marrow transplantation. After a 15-day stay in the transplantation unit, fever developed and imipenem and amikacin were administered. Despite this treatment, the patient died of septic shock syndrome 5 days later. Three-day-old blood cultures grew a carbapenem-resistant P. aeruginosa isolate. This P. aeruginosa COL-1 isolate was resistant to most ß-lactams, including piperacillin/tazobactam, imipenem, meropenem, ceftazidime, cefepime (minimum inhibitory concentrations [MICs] of 128, 32, 16, 64, 32 mg/L, respectively), amikacin, tobramycin, gentamicin, netilmicin, and ciprofloxacin; however, the isolate was susceptible to aztreonam (MIC determination, genetic techniques and ß-lactamase assays are described elsewhere ). A sonicate of crude extract of P. aeruginosa COL-1 culture showed strong imipenem and meropenem hydrolysis activity (0.7 mU/mg and 1.9 mU/mg; reference P. aeruginosa strain <0.05 mU/mg) by UV spectrophotometry with 0.1 mM of substrate, after incubation in 50 mM phosphate buffer at 30°C. This activity was lost when the enzyme extract was preincubated with 10 mM of edetic acid and was partially restored by addition of 1 mM ZnCl2, indicating the presence of a metallo-carbapenem hydrolyzing ß-lactamase. Isoelectric focusing revealed two ß-lactamase bands of pI 5.6 and 9. Only the pI 5.6 ß-lactamase band was inhibited if the gel was overlaid with edetic acid before nitrocefin was added as the indicator substrate; the other pI 9 ß-lactamase likely corresponded to a naturally occurring AmpC cephalosporinase. This pI 5.6 value differed from the pI values of the carbapenem-hydrolyzing ß-lactamase previously reported in P. aeruginosa (3-5,7,8). Polymerase chain reaction amplification experiments were negative when internal primers were used for the only sequenced carbapenem-hydrolyzing ß-lactamase genes from P. aeruginosa encoding IMP-1 and VIM-1 and genomic DNA of P. aeruginosa COL-1. Transfer of the carbapenem resistance marker by conjugation to laboratory strains of P. aeruginosa or Escherichia coli was unsuccessful (9), but transformation by electroporation of a putative plasmid extract from P. aeruginosa COL-1 in E. coli, followed by selection onto amoxicillin-containing agar plates (9), gave a ca. 45-kb plasmid that produced the carbapenem-hydrolyzing ß-lactamase with a pI value of 5.6. Thus, the carbapenem-hydrolyzing ß-lactamase gene was plasmid-borne.
This case indicates the presence of a novel carbapenem-hydrolyzing ß-lactamase in P. aeruginosa in Europe, the first in France; its spread in gram-negative rods, as reported for IMP-1 in Japan, is of concern because, as seen in this case, routine laboratory detection is difficult and therapeutic options are extremely limited.
This work was supported by a grant from the Ministère de l'Education Nationale, de la Recherche et de la Technologie, Université Paris XI, Faculté de Médecine Paris Sud, UPRES-JE-2227, France.
- Livermore DM. Are all ß-lactams created equal? Scand J Infect Dis. 1996;Suppl 101S:33–43.
- Nordmann P, Guibert M. Extended-spectrum ß-lactamases in Pseudomonas aeruginosa. J Antimicrob Chemother. 1998;42:128–31.
- Osano E, Arakawa Y, Wacharotayankun R, Ohta M, Horii T, Ito H, Molecular characterization of an enterobacterial metallo ß-lactamase found in a clinical isolate of Serratia marcescens that shows imipenem resistance. Antimicrob Agents Chemother. 1994;38:71–8.
- Senda K, Arakawa Y, Nakashima K, Ito H, Ichiyama S, Shimokata K, Multifocal outbreaks of metallo-ß-lactamase-producing Pseudomonas aeruginosa resistant to broad-spectrum ß-lactams, including carbapenems. Antimicrob Agents Chemother. 1996;40:349–53.
- Lauretti L, Riccio ML, Mazzariol A, Cornaglia G, Amicosante G, Fontana R, Cloning and characterization of blaVIM, a new integron-borne metallo-ß-lactamase gene from a Pseudomonas aeruginosa clinical isolate. Antimicrob Agents Chemother. 1999;43:1584–90.
- Senda K, Arakawa Y, Ichiyama S, Nakashima K, Ito H, Shimokata K, Characterization of metallo ß-lactamase gene (blaIMP) in gram-negative rods resistant to broad-spectrum ß-lactams. J Clin Microbiol. 1996;34:2909–13.
- Woodford N, Palepou MFI, Babini GS, Bates J, Livermore DM. Carbapenemase-producing Pseudomonas aeruginosa in UK. Lancet. 1998;352:546–7.
- Cardoso O, Sousa JC, Leitao R, Peixe L. Carbapenem-hydrolysing ß-lactamase from clinical isolates of Pseudomonas aeruginosa in Portugal. J Antimicrob Chemother. 1999;44:135.
- Philippon LN, Naas T, Bouthors AT, Barakett V, Nordmann P. OXA-18, a class D clavulanic acid-inhibited extended-spectrum ß-lactamase from Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1997;41:2188–95.
The conclusions, findings, and opinions expressed by authors contributing to this journal do not necessarily reflect the official position of the U.S. Department of Health and Human Services, the Public Health Service, the Centers for Disease Control and Prevention, or the authors' affiliated institutions. Use of trade names is for identification only and does not imply endorsement by any of the groups named above.
- Page created: May 09, 2011
- Page last updated: May 09, 2011
- Page last reviewed: May 09, 2011
- Centers for Disease Control and Prevention,
National Center for Emerging and Zoonotic Infectious Diseases (NCEZID)
Office of the Director (OD)